Regulation of the Gli protein TRA-1 by co-factors

辅因子对 Gli 蛋白 TRA-1 的调节

基本信息

批准号：
9237835
负责人：
RONALD E ELLIS
金额：
$ 30.59万
依托单位：
ROWAN UNIVERSITY SCHOOL/OSTEOPATHIC MED
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-01-01 至 2020-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9237835
关键词：
Activator Appliances Adult Affect Anatomy Animals Anthelmintics Binding Proteins Biological Assay Caenorhabditis Caenorhabditis elegans Cell Fate Control Chromatin Chromatin Structure Cleaved cell Complex DNA Development Developmental Biology Drosophila genus Epigenetic Process Equilibrium Factor Analysis Fertility Fogs Future GLI Family Protein Gene Targeting Genes Genetic Epistasis Genetic Transcription Genetic screening method Germ Lines Human Human Development Lead Learning Malignant Neoplasms Mammals Mass Spectrum Analysis Measures Missense Mutation Modeling Modification Molecular Genetics Mutation Nematoda Orthologous Gene Output Parasitic nematode Pathway interactions Phenotype Play Positioning Attribute Process Protein Isoforms Proteins RNA Interference Regulation Rest Role Spermatogenesis System Textbooks Tissues Transcriptional Regulation Work c new developmental disease experimental study hedgehog signal transduction human disease mutant nematode genetics novel prevent promoter sex determination smoothened signaling pathway transcription factor

项目摘要

Gli proteins constitute a vital but complex group of transcription factors. Humans have three, and mutations in them or errors in their regulation can cause developmental disorders or cancer. Our understanding of these proteins has rested on studies of their Drosophila ortholog Cubitus interruptus, which revealed that Gli proteins are a major target of the Hedgehog-signaling pathway, and can be processed to produce either activators or repressors. However, key questions about Gli function remain unanswered. In particular, it is not clear how much of their activity is controlled by co-factors, or what role they play in epigenetic changes. Since Gli proteins are broad transcriptional regulators, these questions are critical. Caenorhabditis nematodes provide an ideal model for answering these questions. They have a single Gli protein, TRA-1, that shares many similarities with human Gli proteins. However, TRA-1 controls sexual fates and plays a central role in self-fertility. As a result, the balance between its activating and repressing functions in the germ line makes C. briggsae ideal for identifying and evaluating co-factors. Finally, the anatomy of C. briggsae and C. elegans are simple and completely defined, which makes it feasible to study tissue-specific effects of chromatin regulation during development. Similar studies are difficult in other animals. The power of nematode genetics and developmental biology will simply the identification of Gli co-factors, and the analysis of how they work with TRA-1 to activate or repress targets. Indeed, direct transcriptional control and epigenetic effects can both be studied in living animals. Thus, this proposal has three aims: Aim #1: Define TRA-1 activator and repressor functions in Caenorhabditis nematodes. Aim #2: Determine how known co-factors interact with TRA-1 isoforms to regulate target genes. Aim #3: Identify new TRA-1 co-factors. Since TRA-1 is a model Gli protein, many of its co-factors and regulatory interactions are likely to be conserved. Thus, identifying co-factors and elucidating how they work with TRA-1 should illuminate human development and disease. In addition, novel co-factors that are not shared with humans could define new targets for antihelminthic drugs. Since preventing adult worms from reproducing in their human hosts is a critical for managing parasitic nematodes, the ability to target this part of the sex determination pathway could be invaluable. Finally, nematodes are one of the top models for sex determination (as shown by coverage in textbooks like Developmental Biology), so it is critical that we decipher the central part of the story.

Gli 蛋白构成了一组重要但复杂的转录因子。人类有这三种基因，它们的突变或调控错误可能会导致发育障碍或癌症。我们对这些蛋白质的理解依赖于对其果蝇直系同源物 Cubitus Interruptus 的研究，该研究揭示了 Gli 蛋白是 Hedgehog 信号通路的主要靶标，并且可以被加工以产生激活剂或阻遏物。然而，有关 Gli 功能的关键问题仍未得到解答。特别是，尚不清楚它们的活性有多少是由辅助因子控制的，或者它们在表观遗传变化中发挥什么作用。由于 Gli 蛋白是广泛的转录调节因子，这些问题至关重要。秀丽隐杆线虫为回答这些问题提供了理想的模型。它们有一种 Gli 蛋白 TRA-1，与人类 Gli 蛋白有许多相似之处。然而，TRA-1 控制着性命运，并在自体生育方面发挥着核心作用。因此，种系中激活和抑制功能之间的平衡使得 C. briggsae 成为识别和评估辅助因子的理想选择。最后，C. briggsae 和 C. elegans 的解剖结构简单且完全明确，这使得研究发育过程中染色质调节的组织特异性效应成为可能。在其他动物中进行类似的研究很困难。线虫遗传学和发育生物学的力量将简单地识别 Gli 辅助因子，并分析它们如何与 TRA-1 一起激活或抑制靶标。事实上，直接转录控制和表观遗传效应都可以在活体动物中进行研究。因此，该提案具有三个目标：目标#1：定义秀丽隐杆线虫中 TRA-1 激活子和阻遏子功能。目标#2：确定已知辅助因子如何与 TRA-1 同工型相互作用以调节靶基因。目标#3：确定新的 TRA-1 辅助因子。由于 TRA-1 是一种模型 Gli 蛋白，因此它的许多辅助因子和调控相互作用可能是保守的。因此，识别辅助因子并阐明它们如何与 TRA-1 协同作用应该能够阐明人类发育和疾病。此外，与人类不共有的新辅助因子可以定义抗蠕虫药物的新靶点。由于防止成虫在人类宿主中繁殖对于控制寄生线虫至关重要，因此针对性别决定途径的这一部分的能力可能是无价的。最后，线虫是性别决定的顶级模型之一（正如发育生物学等教科书的报道所示），因此我们破译故事的核心部分至关重要。